State Key Laboratory of Animal Nutrition and Feeding, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
J Agric Food Chem. 2024 Feb 28;72(8):3833-3845. doi: 10.1021/acs.jafc.3c07554. Epub 2024 Jan 29.
The acquisition of a thermostable enzyme is an indispensable prerequisite for its successful implementation in industrial applications and the development of novel functionalities. Various protein engineering approaches, including rational design, semirational design, and directed evolution, have been employed to enhance thermostability. However, all of these approaches require sensitive and reliable high-throughput screening (HTS) technologies to efficiently and rapidly identify variants with improved properties. While numerous reviews focus on modification strategies for enhancing enzyme thermostability, there is a dearth of literature reviewing HTS methods specifically aimed at this objective. Herein, we present a comprehensive overview of various HTS methods utilized for modifying enzyme thermostability across different screening platforms. Additionally, we highlight significant recent examples that demonstrate the successful application of these methods. Furthermore, we address the technical challenges associated with HTS technologies used for screening thermostable enzyme variants and discuss valuable perspectives to promote further advancements in this field. This review serves as an authoritative reference source offering theoretical support for selecting appropriate screening strategies tailored to specific enzymes with the aim of improving their thermostability.
获得热稳定性酶是其成功应用于工业应用和开发新功能的必要前提。已经采用了各种蛋白质工程方法，包括合理设计、半理性设计和定向进化，以提高热稳定性。然而，所有这些方法都需要灵敏和可靠的高通量筛选 (HTS) 技术，以便有效地和快速地鉴定具有改进性能的变体。虽然有许多评论集中在增强酶热稳定性的修饰策略上，但很少有文献专门针对这一目标审查 HTS 方法。在这里，我们全面概述了用于在不同筛选平台上修饰酶热稳定性的各种 HTS 方法。此外，我们还强调了一些重要的最新示例，这些示例展示了这些方法的成功应用。此外，我们还讨论了与筛选热稳定酶变体相关的 HTS 技术的技术挑战，并讨论了有价值的观点，以促进该领域的进一步发展。本综述是一个权威的参考来源，为选择适合特定酶的适当筛选策略提供了理论支持，目的是提高其热稳定性。
